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. Author manuscript; available in PMC: 2020 May 6.
Published in final edited form as: Sci Signal. 2015 Nov 3;8(401):ra109. doi: 10.1126/scisignal.aac5356

Fig.5. Truncation of the cytoplasmatic tail results in increased surface expression of iRhom2.

Fig.5.

(A) Immunoprecipitation (IP) for T7 or ADAM17 followed by immunoblotting for the same in lysates from L-929 cells stably expressing T7-tagged WT or truncated iRhom2. Blot is representative of three experiments. (B) Immunoblotting for iRhom2 using a T7 antibody in intracellular and cell surface fractions from L-929 cells stably expressing WT or truncated iRhom2. Blot is representative of three experiments. (C) Surface abundance of iRhom2, determined using an antibody against T7, on stably transfected L-929 cells (n = 8). (D) Immunocytochemistry for iRhom2 using T7 antibodies (Cy3), phalloidin–fluorescein isothiocyanate (FITC), and Hoechst staining in stably transfected L-929 cells, fixed, and/or permeabilized as indicated (n ≥ 3 experiments). (E and F) Flow cytometry analysis of MFI of the surface abundance of iRhom2 on unpermeabilized stably transfected L-929 cells expressing either scrambled or ADAM17 shRNA (n ≥ 4 experiments) (E) or immortalized WT or Adam17 knockout (KO) mouse embryonic fibroblasts (MEFs) (n = 12 experiments) (F). Data are means ± SEM from the number of experiments (n) indicated; ***P < 0.001. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. Na+/K+ ATPase, Na+- and K+-dependent adenosine triphosphatase.